LOCUS Exported 6468 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION Booster-ERK, a Red-Shifted Genetically Encoded FRET Biosensor for ERK activity.. ACCESSION . VERSION . KEYWORDS pCAGGS-4511NES SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 6468) AUTHORS Watabe T, Terai K, Sumiyama K, Matsuda M TITLE Booster, a Red-Shifted Genetically Encoded Forster Resonance Energy Transfer (FRET) Biosensor Compatible with Cyan Fluorescent Protein/Yellow Fluorescent Protein-Based FRET Biosensors and Blue Light-Responsive Optogenetic Tools. JOURNAL ACS Sens. 2020 Feb 26. doi: 10.1021/acssensors.9b01941. PUBMED 32101394 REFERENCE 2 (bases 1 to 6468) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..6468 /organism="synthetic DNA construct" /mol_type="other DNA" enhancer 1..380 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" primer_bind 1536..1558 /label=pCAGGS-5 /note="Chimeric intron in CAG promoter, forward primer" primer_bind 1624..1643 /label=pCAG-F /note="Rabbit beta-globin intron, for pCAG plasmids, forward primer" regulatory 1673..1682 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" CDS 1679..2374 /codon_start=1 /product="monomeric far-red fluorescent protein (Shcherbo et al., 2009)" /label=mKate2 /note="mammalian codon-optimized" /translation="MVSELIKENMHMKLYMEGTVNNHHFKCTSEGEGKPYEGTQTMRIK AVEGGPLPFAFDILATSFMYGSKTFINHTQGIPDFFKQSFPEGFTWERVTTYEDGGVLT ATQDTSLQDGCLIYNVKIRGVNFPSNGPVMQKKTLGWEASTETLYPADGGLEGRADMAL KLVGGGHLICNLKTTYRSKKPAKNLKMPGVYYVDRRLERIKEADKETYVEQHEVAVARY CDLPSKLGHR" CDS 2711..3364 /codon_start=1 /product="monomeric Kusabira-Orange 2 fluorescent protein" /label=mKO2 /translation="MVSVIKPEMKMRYYMDGSVNGHEFTIEGEGTGRPYEGHQEMTLRV TMAEGGPMPFAFDLVSHVFCYGHRVFTKYPEEIPDYFKQAFPEGLSWERSLEFEDGGSA SVSAHISLRGNTFYHKSKFTGVNFPADGPIMQNQSVDWEPSTEKITASDGVLKGDVTMY LKLEGGGNHKCQFKTTYKAAKEILEMPGDHYIGHRLVRKTEGNITEQVEDAVAHS" CDS 3440..3466 /codon_start=1 /product="nuclear export signal from the HIV Rev protein (Fischer et al., 1995)" /label=NES /translation="LPPLERLTL" primer_bind complement(3482..3501) /label=Bglob-pA-R /note="Rabbit beta-globin polyA region, reverse primer" polyA_signal 3547..3602 /label=-beta-globin poly(A) signal /note="rabbit beta-globin polyadenylation signal (Gil and Proudfoot, 1987)" primer_bind complement(3601..3620) /label=rbglobpA-R /note="Rabbit beta-globin polyA, reverse primer. Also called rb-glob-pA-term-R" primer_bind complement(3963..3979) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(3963..3979) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(3976..3998) /label=M13/pUC Reverse /note="In lacZ gene" protein_bind 3987..4003 /label=lac operator /bound_moiety="lac repressor encoded by lacI" /note="The lac repressor binds to the lac operator to inhibit transcription in E. coli. This inhibition can be relieved by adding lactose or isopropyl-beta-D-thiogalactopyranoside (IPTG)." promoter complement(4011..4041) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 4056..4077 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." promoter 4136..4331 /label=SV40 promoter /note="SV40 early promoter" rep_origin 4182..4317 /label=SV40 ori /note="SV40 origin of replication" primer_bind 4244..4263 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" polyA_signal 4337..4471 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(4374..4393) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 4428..4447 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" primer_bind complement(4539..4556) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(4710..5298) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(4790..4809) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(5469..6329) /codon_start=1 /gene="bla" /product="beta-lactamase" /label=AmpR /note="confers resistance to ampicillin, carbenicillin, and related antibiotics" /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS LIKHW" primer_bind 6092..6111 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(6330..6434) /gene="bla" /label=AmpR promoter ORIGIN 1 gacattgatt attgactagt tattaatagt aatcaattac ggggtcatta gttcatagcc 61 catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca 121 acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga 181 ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc 241 aagtgtatca tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct 301 ggcattatgc ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat 361 tagtcatcgc tattaccatg gtcgaggtga gccccacgtt ctgcttcact ctccccatct 421 cccccccctc cccaccccca attttgtatt tatttatttt ttaattattt tgtgcagcga 481 tgggggcggg gggggtgcgg cggcagccaa tcagagcggc gcgctccgaa agtttccttt 541 tatggcgagg cggcggcggc ggcggcccta taaaaagcga agcgcgcggc gggcgggagt 601 cgctgcgcgc tgccttcgcc ccgtgccccg ctccgccgcc gcctcgcgcc gcccgccccg 661 gctctgactg accgcgttac tcccacaggt gagcgggcgg gacggccctt ctcctccggg 721 ctgtaattag cgcttggttt aatgacggct tgtttctttt ctgtggctgc gtgaaagcct 781 tgaggggctc cgggagggcc ctttgtgcgg ggggagcggc tcggggggtg cgtgcgtgtg 841 tgtgtgcgtg gggagcgccg cgtgcggctc cgcgctgccc ggcggctgtg agcgctgcgg 901 gcgcggcgcg gggctttgtg cgctccgcag tgtgcgcgag gggagcgcgg ccgggggcgg 961 tgccccgcgg tgcggggggg gctgcgaggg gaacaaaggc tgcgtgcggg gtgtgtgcgt 1021 gggggggtga gcagggggtg tgggcgcgtc ggtcgggctg caaccccccc tgcacccccc 1081 tccccgagtt gctgagcacg gcccggcttc gggtgcgggg ctccgtacgg ggcgtggcgc 1141 ggggctcgcc gtgccgggcg gggggtggcg gcaggtgggg gtgccgggcg gggcggggcc 1201 gcctcgggcc ggggagggct cgggggaggg gcgcggcggc ccccggagcg ccggcggctg 1261 tcgaggcgcg gcgagccgca gccattgcct tttatggtaa tcgtgcgaga gggcgcaggg 1321 acttcctttg tcccaaatct gtgcggagcc gaaatctggg aggcgccgcc gcaccccctc 1381 tagcgggcgc ggggcgaagc ggtgcggcgc cggcaggaag gaaatgggcg gggagggcct 1441 tcgtgcgtcg ccgcgccgcc gtccccttct ccctctccag cctcggggct gtccgcgggg 1501 ggacggctgc cttcgggggg gacggggcag ggcggggttc ggcttctggc gtgtgaccgg 1561 cggctctagc tagagcctct gctaaccatg ttcatgcctt cttctttttc ctacagctcc 1621 tgggcaacgt gctggttatt gtgctgtctc atcattttgg caagaattcg ccgccaccat 1681 ggtgagcgag ctgattaagg agaacatgca catgaagctg tacatggagg gcaccgtgaa 1741 caaccaccac ttcaagtgca catccgaggg cgaaggcaag ccctacgagg gcacccagac 1801 catgagaatc aaggcggtcg agggcggccc tctccccttc gccttcgaca tcctggctac 1861 cagcttcatg tacggcagca aaaccttcat caaccacacc cagggcatcc ccgacttctt 1921 taagcagtcc ttccccgagg gcttcacatg ggagagagtc accacatacg aagacggggg 1981 cgtgctgacc gctacccagg acaccagcct ccaggacggc tgcctcatct acaacgtcaa 2041 gatcagaggg gtgaacttcc catccaacgg ccctgtgatg cagaagaaaa cactcggctg 2101 ggaggcctcc accgagaccc tgtaccccgc tgacggcggc ctggaaggca gagccgacat 2161 ggccctgaag ctcgtgggcg ggggccacct gatctgcaac ttgaagacca catacagatc 2221 caagaaaccc gctaagaacc tcaagatgcc cggcgtctac tatgtggaca gaagactgga 2281 aagaatcaag gaggccgaca aagagaccta cgtcgagcag cacgaggtgg ctgtggccag 2341 atactgcgac ctccctagca aactggggca cagactcgag atggcggacg aggagaagct 2401 gccgcccggc tgggagaagc gcatgagccg cagctcaggc cgagtgtact acttcaacca 2461 catcactaac gccagccagt gggagcggcc cagcggcaac agcagcagtg gtggcaaaaa 2521 cgggcagggg gagcctgcca ggggtaccag tgctggtggt agtgctggtg gtagtgctgg 2581 tggtagtgct ggtggtagtg ctggtggttc cggcagtgct ggtggtagtg ctggtggtag 2641 taccagtgct ggtggtagtg ctggtggtag tgctggtggt agtgctggtg gtagtgctgg 2701 tggttccgga atggtgagtg tgattaaacc agagatgaag atgaggtact acatggacgg 2761 ctccgtcaat gggcatgagt tcacaattga aggtgaaggc acaggcagac cttacgaggg 2821 acatcaagag atgacactac gcgtcacaat ggccgagggc gggccaatgc ctttcgcgtt 2881 tgacttagtg tcacacgtgt tctgttacgg ccacagagta tttactaaat atccagaaga 2941 gataccagac tatttcaaac aagcatttcc tgaaggcctg tcatgggaaa ggtcgttgga 3001 gttcgaagat ggtgggtccg cttcagtcag tgcgcatata agccttagag gaaacacctt 3061 ctaccacaaa tccaaattta ctggggttaa ctttcctgcc gatggtccta tcatgcaaaa 3121 ccaaagtgtt gattgggagc catcaaccga gaaaattact gccagcgacg gagttctgaa 3181 gggtgatgtt acgatgtacc taaaacttga aggaggcggc aatcacaaat gccaattcaa 3241 gactacttac aaggcggcaa aagagattct tgaaatgcca ggagaccatt acatcggcca 3301 tcgcctcgtc aggaaaaccg aaggcaacat tactgagcag gtagaagatg cagtagctca 3361 ttccgcggcc gcaccagacg tccctagaac accggtggat aaagcaaagc tgtcattcca 3421 atttccgtct agactgcagc tgcctcctct ggaacgcctg actctggatt aagtcgacgg 3481 atctttttcc ctctgccaaa aattatgggg acatcatgaa gccccttgag catctgactt 3541 ctggctaata aaggaaattt attttcattg caatagtgtg ttggaatttt ttgtgtctct 3601 cactcggaag gacatatggg agggcaaatc atttaaaaca tcagaatgag tatttggttt 3661 agagtttggc aacatatgcc catatgctgg ctgccatgaa caaaggttgg ctataaagag 3721 gtcatcagta tatgaaacag ccccctgctg tccattcctt attccataga aaagccttga 3781 cttgaggtta gatttttttt atattttgtt ttgtgttatt tttttcttta acatccctaa 3841 aattttcctt acatgtttta ctagccagat ttttcctcct ctcctgacta ctcccagtca 3901 tagctgtccc tcttctctta tggagatccc tcgacctgca gcccaagctt ggcgtaatca 3961 tggtcatagc tgtttcctgt gtgaaattgt tatccgctca caattccaca caacatacga 4021 gccggaagca taaagtgtaa agcctggggt gcctaatgag tgagctaact cacattaatt 4081 gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt cgtgccagcg gatccgcatc 4141 tcaattagtc agcaaccata gtcccgcccc taactccgcc catcccgccc ctaactccgc 4201 ccagttccgc ccattctccg ccccatggct gactaatttt ttttatttat gcagaggccg 4261 aggccgcctc ggcctctgag ctattccaga agtagtgagg aggctttttt ggaggcctag 4321 gcttttgcaa aaagctaact tgtttattgc agcttataat ggttacaaat aaagcaatag 4381 catcacaaat ttcacaaata aagcattttt ttcactgcat tctagttgtg gtttgtccaa 4441 actcatcaat gtatcttatc atgtctggat ccgctgcatt aatgaatcgg ccaacgcgcg 4501 gggagaggcg gtttgcgtat tgggcgctct tccgcttcct cgctcactga ctcgctgcgc 4561 tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat acggttatcc 4621 acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca aaaggccagg 4681 aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat 4741 cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag 4801 gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga 4861 tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg 4921 tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt 4981 cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac 5041 gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc 5101 ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag aacagtattt 5161 ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc 5221 ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc 5281 agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg 5341 aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag 5401 atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg 5461 tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt 5521 tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatacggga gggcttacca 5581 tctggcccca gtgctgcaat gataccgcga gacccacgct caccggctcc agatttatca 5641 gcaataaacc agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgcc 5701 tccatccagt ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt 5761 ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg 5821 gcttcattca gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtgc 5881 aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg 5941 ttatcactca tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtaaga 6001 tgcttttctg tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga 6061 ccgagttgct cttgcccggc gtcaatacgg gataataccg cgccacatag cagaacttta 6121 aaagtgctca tcattggaaa acgttcttcg gggcgaaaac tctcaaggat cttaccgctg 6181 ttgagatcca gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact 6241 ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata 6301 agggcgacac ggaaatgttg aatactcata ctcttccttt ttcaatatta ttgaagcatt 6361 tatcagggtt attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa 6421 ataggggttc cgcgcacatt tccccgaaaa gtgccacctg ggtcgatc //